Resonant sensors and methods of use thereof for the determination of analytes

• US 8,349,611 B2
• Filed: 02/17/2010
• Issued: 01/08/2013
• Est. Priority Date: 02/17/2009
• Status: Expired due to Fees

• Current Assignee: Leversense LLC
• Original Assignee: Leversense LLC
• Inventors: Loverich, Jacob J., Frank, Jeremy E., Nagy, Peter A.
• Primary Examiner(s): Soderquist, Arlen
• Application Number:
  US12/707,254
• Publication Number:
  US 20100207602A1
• Time in Patent Office: 1,056 Days
• Field of Search: 73/240.1, 73/240.6, 735/79, 422/82.01, 436/86, 436/94, 436/178, 436/512, 436/524, 436/527
• US Class Current: 436/86
• CPC Class Codes: G01N 2291/012 Phase angle
• CPC Class Codes: G01N 2291/0255 (Bio)chemical reactions, e.g. on biosensors
• CPC Class Codes: G01N 2291/0256 Adsorption, desorption, surface mass change, e.g. on biosensors
• CPC Class Codes: G01N 2291/0427 Flexural waves, plate waves, e.g. Lamb waves, tuning fork, cantilever
• CPC Class Codes: G01N 29/022 Fluid sensors based on microsensors, e.g. quartz crystal-microbalance [QCM], surface acoustic wave [SAW] devices, tuning forks, cantilevers, flexural plate wave [FPW] devices microdevices per se B81B
• CPC Class Codes: G01N 29/036 by measuring frequency or resonance of acoustic waves
• CPC Class Codes: G01N 29/2437 Piezoelectric probes
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

Resonant sensors, preferably having floating bilayer symmetry, and their methods of use is determining the presence, amount or binding kinetics of an analyte of interest in a test sample are disclosed. The test sample may be a liquid or gas.

Claims

1. A resonant sensor for determining the presence, amount or binding kinetics of an analyte in a test medium, said sensor being operable under the influence of an activating force field and comprising:

an active, deformable planar member;

a passive, deformable planar member attached to said active, deformable planar member, forming a pliable, layered sensing element having flexural stiffness and a resonant flexural bending or extensional mode for analyte determination and at least one axis of symmetry;

at least one substrate coupled to said layered sensing element by at least one flexible coupling element having a first end joined to said substrate and a second end joined to said sensing element, said substrate having a flexible portion and permitting said vibration of said sensing element;

said layered sensing element being supported by said flexible portion of said substrate to which at least one of the planar members of said sensing element is connected, the flexible substrate portion having a bending modulus which is less than the sensing element bending modulus; and

a capture agent for said analyte associated with at least one exterior surface site of said sensing element, said capture agent being one member of a specific binding pair and the other member of said specific binding pair being said analyte;

said sensing element with associated capture agent being caused to vibrate at a determinable resonance frequency by said activating force field operating on said active, deformable planar member, said activating force field inducing in said sensing element a uniform bending moment or extensional force component that is substantially parallel or perpendicular to said at least one axis of symmetry.

2. The resonant sensor according to claim 1, wherein said passive, deformable planar member is coextensive with said active, deformable planar member.

3. The resonant sensor according to claim 1, wherein said active, deformable planar member comprises a piezoelectric material, and said activating force field is an electric field.

4. The resonant sensor according to claim 1, wherein said active, deformable planar member comprises an electrostrictive material, and said activating force field is an electric field.

5. The resonant sensor according to claim 1, wherein said active, deformable planar member comprises a magnetostrictive material, and said activating force field is a magnetic field.

6. The resonant sensor according to claim 1 having flexural stiffness about the axis of symmetry of said sensing element at the junction between the flexible coupling element and the sensing element, which is an order of magnitude less than the flexural stiffness of the sensing element, and the total mass of the flexible coupling element intermediate its ends is an order of magnitude less than the mass of the sensing element.

7. The resonant sensor according to claim 6, wherein said activating force field is provided by an electric voltage source and said coupling element comprises an electrical conduction path from said voltage source to at least one side of said active planar member.

8. The resonant sensor according to claim 1, wherein both the active and passive planar members of said sensing element are embedded in the flexible portion of said at least one substrate.

9. The resonant sensor according to claim 1, wherein the in-plane shape of the sensing element is selected from the group consisting of a circular, square or rectangular shape.

10. The resonant sensor according to claim 9, wherein the in-plane shape of the sensing element is rectangular and the length dimension of said active planar member is greater than the length dimension of said passive planar member.

11. The resonant sensor according to claim 9, wherein the in-plane shape of the sensing element is rectangular and the length dimension of said passive planar member is greater than the length dimension of the active planar member.

12. The resonant sensor according to claim 10 or 11, wherein the flexural bending or extensional mode used for analyte determination is selected from (i) the first vibration bending or extending mode along the width dimension of the sensing element, or (ii) an integer multiple of said first vibration bending mode along said width dimension.

13. The resonant sensor according to claim 9, wherein the in-plane shape of the sensing element is circular and the total thickness of the planar members constituting said sensing element is less than the radial dimension of said sensing element.

14. The resonant sensor according to claim 13, wherein the flexural bending or extensional mode used for analyte determination is selected from (i) the first membrane bending mode along the radial dimension of said sensing element, or (ii) an integer multiple of said first membrane bending mode along said radial dimension.

15. The resonant sensor according to claim 13, wherein the thickness of said layered sensing element is from about 0.05 mm to about 5 mm and the radial dimension is from about 0.25 mm to about 10 mm.

16. The resonant sensor according to claim 9, wherein the in-plane shape of the sensing element is rectangular or square and the total thickness of the planar members constituting said sensing element is less than the in-plane length or width dimension of said sensing element.

17. The resonant sensor according to claim 16, wherein the flexural bending or extensional mode used for analyte determination is selected from (i) the first vibration bending or extending mode along the length dimension of said sensing element or (ii) an integer multiple of said first vibration bending mode along said length dimension.

18. The resonant sensor according to claim 16, wherein the thickness of said layered sensing element is from about 0.05 mm to about 5 mm and the length and width dimensions are from about 0.25 mm to about 10 mm.

19. The resonant sensor according to claim 18, wherein the dimensions of the layered sensing element are 0.250 mm thick, 1.0 mm wide and 4.0 mm long.

20. The resonant sensor according to claim 18, wherein the thickness of the passive, deformable planar member is about 0.010 um to about 250 um.

21. The resonant sensor according to claim 18, wherein the passive, deformable planar member is in the form of a thin electrode on the active, deformable planar member.

22. The resonant sensor according to claim 1, wherein said in-plane shape of the sensing element is rectangular and said capture agent is disposed on the exterior surface of said element at sites spaced apart from nodal points or lines that are characteristic of said flexural bending or extensional mode.

23. The resonant sensor according to claim 1, wherein said activating force activates the active, deformable planar member, causing portions of the sensing element adjacent to nodal points or lines to flex out of the plane or extend in plane of the sensing element, said nodal points or lines being characteristic of said flexural bending or extensional mode.

24. An apparatus for determining the presence, or amount or binding kinetics of an analyte in a test medium, said apparatus, comprising a resonant sensor as claimed in claim 1, and a device operable to determine said resonance frequency.

25. A system for determining the presence, or amount or binding kinetics of an analyte in a test medium, said system, comprising an assembly of concatenated resonance sensors, as claimed in claim 1, and a device operable to determine the resonance frequency of the resonant sensors in said assembly.

26. The system according to claim 25, wherein each of the concatenated sensors has a different dynamic range.

27. The system according to claim 25, wherein each of the resonant sensors in said assembly is operable under the influence of a common activating force field, and the resonance frequency at which the sensing element of each said resonance sensor vibrates is determined by a common frequency determining device.

28. The system according to claim 25, wherein each of the resonant sensors in said assembly has disposed on the surface thereof a capture agent which is different from the capture agent on any other sensing element in said assembly, whereby each resonant sensor captures a different analyte.

29. The resonant sensor according to claim 1, wherein said specific binding pair member and the analyte captured by said specific binding pair member form a binding pair selected from the group consisting of nucleic acid (RNA or DNA) fragments-complementary sequences, antigen-antibody, receptor-hormone, receptor-ligand, agonist-antagonist, avidin-biotin, virus-receptor.

30. The resonant sensor according to claim 29, wherein said binding pair comprises nucleic acid fragment-complementary sequence.

31. The resonant sensor according to claim 30, wherein one binding pair member is selected from the group of DNA molecules, RNA molecules, cDNA molecules, cRNA molecules, RNAi molecules and pRNA molecules.

32. The resonant sensor according to claim 1, wherein the at least one flexible coupling element is critically damped at the resonance frequencies used for analyte determination.

33. The resonant sensor according to claim 1 wherein the flexural stiffness about the axis of symmetry of said sensing element at the junction between said flexible portion(s) and the sensing element is an order of magnitude less than the flexural stiffness of the sensing element, and the total mass of said flexible portion(s) is an order of magnitude less than the mass of the sensing element.

34. A method for determining an analyte of interest in a test medium suspected to contain said analyte, which is a member of a specific binding pair, said method comprising:

a. providing at least one resonant sensor as claimed in claim 1;

b. establishing a reference resonance frequency;

c. contacting said sensor with a medium suspected to contain an analyte of interest, for a time sufficient for said specific binding pair member to bind analyte of interest present in said medium;

d. determining a test resonance frequency of said sensor after contact with said medium, a shift between the test resonance frequency and the reference resonance frequency providing a qualitative indication of target analyte in said test medium.

35. The method of claim 34 further comprising quantitatively determining the analyte of interest in said test medium by measuring said frequency shift associated with analyte binding, the difference in frequency being related to the amount/mass of the bound analyte.

36. The method of claim 34 or 35, wherein said capture agent comprises a single stranded nucleic acid molecule comprised of a series of nucleotide(s) and the test medium contacted with said resonant sensor is suspected of containing a target nucleic acid analyte comprising a nucleotide sequence complementary to said single stranded nucleic acid.

37. The method of claim 36, further comprising exposing said resonant sensor to polymerase activity.

38. The method of claim 34, wherein said nucleic acid molecule is selected from the group of DNA molecules, RNA molecules, cDNA molecules, cRNA molecules, RNAi molecules and pRNA molecules.

39. A resonant sensor for determining the presence, amount or binding kinetics of an analyte in a test medium, said sensor being operable under the influence of an activating force field and comprising:

an active, deformable planar member, comprising a piezoelectric material and having first and second metallized surfaces;

a passive, deformable planar member, attached to a metalized surface of said active deformable planar member, forming a pliable, layered sensing element having flexural stiffness and a resonant flexural bending or extensional mode for analyte determination and at least one axis of symmetry;

at least one substrate coupled to said layered sensing element by at least one flexible coupling element having a first end joined to said substrate and a second end joined to said sensing element, said substrate supporting and permitting said vibration of said sensing element; and

a capture agent for said analyte associated with at least one exterior surface site of said sensing element;

said sensing element with associated capture agent being caused to vibrate at a determinable resonance frequency by said activating force field operating on said active, deformable planar member, said activating force field inducing in said sensing element a uniform bending moment or extensional force component that is substantially parallel or perpendicular to said at least one axis of symmetry, said activating force field being an electric field.

40. The resonant sensor according to claim 39, wherein said capture agent for said analyte is an absorbent material effective to absorb said analyte.

41. The resonant sensor according to claim 40, wherein said absorbent material is a polymeric absorbent.